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Plasmid Files

YEp24

Yeast episomal cloning vector with a URA3 marker.

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YEp24 Sequence and MapYEp24.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 ZraI (7697) AseI (6949) AhdI (6777) PflFI - Tth111I (5631) PvuII (5477) BsaBI * (5081) BspEI * (5073) EagI (4348) PshAI (4125) SalI (4060) EcoNI (4035) SphI (3975) AatII (7699) BclI * (304) MfeI (390) XbaI (703) SnaBI (1044) HpaI (1686) BlpI (2125) BspDI - ClaI (2267) SbfI (2481) NcoI (2704) PspOMI * (2873) ApaI * (2877) TspMI - XmaI (3378) SmaI (3380) NheI (3638) BmtI (3642) BamHI (3784) SgrAI (3819) YEp24 7769 bp
ZraI  (7697)
1 site
G A C G T C C T G C A G
AseI  (6949)
1 site
A T T A A T T A A T T A
AhdI  (6777)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
PflFI  (5631)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (5631)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PvuII  (5477)
1 site
C A G C T G G T C G A C
BsaBI  (5081)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BspEI  (5073)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
EagI  (4348)
1 site
C G G C C G G C C G G C
PshAI  (4125)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SalI  (4060)
1 site
G T C G A C C A G C T G
EcoNI  (4035)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
SphI  (3975)
1 site
G C A T G C C G T A C G
AatII  (7699)
1 site
G A C G T C C T G C A G
BclI  (304)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MfeI  (390)
1 site
C A A T T G G T T A A C
XbaI  (703)
1 site
T C T A G A A G A T C T
SnaBI  (1044)
1 site
T A C G T A A T G C A T
HpaI  (1686)
1 site
G T T A A C C A A T T G
BlpI  (2125)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BspDI  (2267)
1 site
A T C G A T T A G C T A
ClaI  (2267)
1 site
A T C G A T T A G C T A
SbfI  (2481)
1 site
C C T G C A G G G G A C G T C C
NcoI  (2704)
1 site
C C A T G G G G T A C C
PspOMI  (2873)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI  (2877)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI can be used between 25°C and 37°C.
TspMI  (3378)
1 site
C C C G G G G G G C C C
XmaI  (3378)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (3380)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
NheI  (3638)
1 site
G C T A G C C G A T C G
BmtI  (3642)
1 site
G C T A G C C G A T C G
BamHI  (3784)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
SgrAI  (3819)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
2μ ori
344 .. 1686  =  1343 bp
yeast 2μ plasmid origin of replication
2μ ori
344 .. 1686  =  1343 bp
yeast 2μ plasmid origin of replication
TcR
3495 .. 4685  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
3495 .. 4685  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
AmpR
6704 .. 7564  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   6704 .. 7495  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6704 .. 7564  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   7496 .. 7564  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6704 .. 7564  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
URA3
2499 .. 3302  =  804 bp
267 amino acids  =  29.3 kDa
Product: orotidine-5'-phosphate decarboxylase,
required for uracil biosynthesis
yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)
URA3
2499 .. 3302  =  804 bp
267 amino acids  =  29.3 kDa
Product: orotidine-5'-phosphate decarboxylase,
required for uracil biosynthesis
yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)
ori
5945 .. 6533  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
5945 .. 6533  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
URA3 promoter
2283 .. 2498  =  216 bp
URA3 promoter
2283 .. 2498  =  216 bp
rop
5326 .. 5517  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
5326 .. 5517  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
AmpR promoter
7565 .. 7669  =  105 bp
AmpR promoter
7565 .. 7669  =  105 bp
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